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Development of a novel immunobiosensor method for the rapid detection of okadaic acid contamination in shellfish extracts

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Abstract

The mouse bioassay is the methodology that is most widely used to detect okadaic acid (OA) in shellfish samples. This is one of the best-known toxins, and it belongs to the family of marine biotoxins referred to as the diarrhetic shellfish poisons (DSP). Due to animal welfare concerns, alternative methods of toxin detection are being sought. A rapid and specific biosensor immunoassay method was developed and validated for the detection of OA. An optical sensor instrument based on the surface plasmon resonance (SPR) phenomenon was utilised. A polyclonal antibody to OA was raised against OA–bovine thyroglobulin conjugate and OA–N-hydroxy succinimide ester was immobilised onto an amine sensor chip surface. The assay parameters selected for the analysis of the samples were: antibody dilution, 1/750; ratio of antibody to standard, 1:1; volume of sample injected, 25 μl min−1; flow rate, 25 μl min−1. An assay action limit of 126 ng g−1 was established by analysing of 20 shellfish samples spiked with OA at the critical concentration of 160 ng g−1, which is the action limit established by the European Union (EU). At this concentration of OA, the assay delivered coefficient of variations (CVs) of <10%. The chip surface developed was shown to be highly stable, allowing more than 50 analyses per channel. When the concentrations of OA determined with the biosensor method were compared with the values obtained by LC–MS in contaminated shellfish samples, the correlation between the two analytical methods was found to be highly satisfactory (r 2 = 0.991).

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Acknowledgements

We thank the European project under the Sixth Framework Programme Priority 5 Food Quality and Safety (Contract No.: Detectox, 514055) for funding.

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Authors and Affiliations

  1. Institute of Agri-Food and Land Use, Queen’s University of Belfast, Stranmillis Road, BT9 5AG, Belfast, Northern Ireland, UK

    Nuria M. Llamas, Linda Stewart & Christopher T. Elliott

  2. Agri-Food and Biosciences Institute, Stoney Road, Stormont, BT4 3SD, Belfast, Northern Ireland, UK

    Terry Fodey & H. Cowan Higgins

  3. Community Reference Laboratory of Marine Biotoxins (CRLMB), Estación Marítima s/n, 36200, Vigo, Spain

    María Luisa R. Velasco & Luis M. Botana

Authors
  1. Nuria M. Llamas
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  2. Linda Stewart
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  3. Terry Fodey
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  4. H. Cowan Higgins
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  5. María Luisa R. Velasco
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  6. Luis M. Botana
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  7. Christopher T. Elliott
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Correspondence to Christopher T. Elliott.

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Llamas, N.M., Stewart, L., Fodey, T. et al. Development of a novel immunobiosensor method for the rapid detection of okadaic acid contamination in shellfish extracts. Anal Bioanal Chem 389, 581–587 (2007). https://doi.org/10.1007/s00216-007-1444-3

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  • DOI: https://doi.org/10.1007/s00216-007-1444-3

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